Andrä S, Frey G, Jaenicke R, Stetter K O
Lehrstuhl für Mikrobiologie, Universität Regensburg, Germany.
Eur J Biochem. 1998 Jul 1;255(1):93-9. doi: 10.1046/j.1432-1327.1998.2550093.x.
The ATPase activity of the thermosome from a methanogen, Methanopyrus kandleri, was characterized in detail. In contrast to all other known chaperonins, enzymatic ATP hydrolysis was found to be strictly dependent on high levels of ammonium salts in vitro. The ths gene encoding the thermosome subunit from the hyperthermophilic M. kandleri was functionally expressed in Escherichia coli and the overproduced polypeptide was assembled into intact thermosome complexes in the mesophilic host. The recombinant particles could be purified by a simple two-step procedure including only one chromatographic step. Structural and biochemical properties of the recombinant protein were closely similar to those of the natural complex. Western blot analysis with an antiserum against the M. kandleri thermosome indicated the presence of at least two subfamilies of archaeal chaperonins.
对来自产甲烷菌坎氏甲烷嗜热菌（Methanopyrus kandleri）的热体的ATP酶活性进行了详细表征。与所有其他已知的伴侣蛋白不同，发现体外酶促ATP水解严格依赖于高浓度的铵盐。编码嗜热栖热菌（Thermus thermophilus）热体亚基的ths基因在大肠杆菌中实现了功能表达，过量产生的多肽在嗜温宿主中组装成完整的热体复合物。重组颗粒可以通过简单的两步程序纯化，仅包括一步色谱步骤。重组蛋白的结构和生化特性与天然复合物非常相似。用抗坎氏甲烷嗜热菌热体的抗血清进行的蛋白质印迹分析表明存在至少两个古菌伴侣蛋白亚家族。
